Structural features of the human bradykinin B2 receptor probed by agonists, antagonists, and anti-idiotypic antibodies

Alla, Said Abd; Buschko, J; Quitterer, Ursula; Maidhof, Armin; Haasemann, M; Breipohl, Gerhard; Knolle, J.; Müller‐Esterl, Werner

doi:10.1016/s0021-9258(19)85333-4

Cited by 66 publications

(7 citation statements)

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“…This ligand, which binds to B 2 receptors with an affinity of K i ) (1.5 ( 0.3) × 10 -9 M, specifically labeled B 2 receptors. The molecular mass of 69 ( 3 kDa (Figure 7A, lanes 1and 2) is in good agreement with previous findings (AbdAlla et al, 1993). The labeling of the B 2 receptor was suppressed when the cross-linking was performed in the presence of a 1000-fold molar excess of bradykinin (Figure 7A, lane 3).…”

Section: Characterization Of B 2 Receptors Endogenously Expressed On ...supporting

confidence: 91%

“…A feature of several 7TM receptors known long before the cDNA sequences of the receptors have been cloned is the reciprocal modulation of agonist binding versus antagonist binding by monovalent cations (Pert & Snyder, 1974; Tsai & Lefkowitz, 1978). Several observations suggest that an aspartate (Horstman et al, 1990) at the intracellular edge of transmembrane domain-II (TM-II) which is highly conserved among G protein-coupled receptors (Baldwin, 1993) is at least in part responsible for this effect.…”

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confidence: 99%

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Na⁺ Ions Binding to the Bradykinin B₂ Receptor Suppress Agonist-Independent Receptor Activation

et al. 1996

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Control of the balance between receptor activation and inactivation is a prerequisite for seven transmembrane domain (7TM) receptor function. We asked for a mechanism to stabilize the inactive receptor conformation which prevents agonist-independent receptor activation. Na+ ions have reciprocal effects on agonist versus antagonist interaction with various 7TM receptors. To investigate the Na+ dependence of receptor activation we chose the bradykinin B2 receptor as a prototypic 7TM receptor. Decrease of the intracellular Na+ content from 40 mM to 10 mM of COS-1 cells transiently expressing rat B2 receptors activated the B2 receptor in the absence of agonist as shown by a 3-fold increase in the basal release of inositolphosphates and increased the intrinsic activity of bradykinin to 1.2. In contrast, under increased intracellular Na+ (148 mM) the intrinsic activity of bradykinin decreased to 0.72. When the interaction of Na+ with the B2 receptor was prevented by exchanging a conserved aspartate in transmembrane domain II for asparagine the B2 receptor was also constitutively-activated in the absence of agonist. Agonist-independence B2 receptor activation under decreased intracellular Na+ was similarly observed with primary human fibroblasts endogenously expressing human B2 receptors by a 2.5-fold increase in basal inositolphosphates. Activation of human B2 receptors in the absence of agonist under decreased intracellular Na+ was further evident by an increased basal phosphorylation of the B2 receptor protein. Thus our data suggest that the interaction of Na+ ions with the B2 receptor stabilizes or induces an inactive receptor conformation thereby providing a mechanism to suppress agonist-independent receptor activation in vivo.

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Section: Characterization Of B 2 Receptors Endogenously Expressed On ...supporting

confidence: 91%

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confidence: 99%

Na⁺ Ions Binding to the Bradykinin B₂ Receptor Suppress Agonist-Independent Receptor Activation

et al. 1996

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“…Materials. The chemiluminescence detection kit and [ 35 S]methionine were from Amersham; [2,3-prolyl-3,4- 3 H]bradykinin (specific activity 98 Ci/mmol) was from NEN Dupont; 1,5-difluoro-2,4-dinitrobenzene (DFDNB) was from Pierce; wheat germ agglutinin (WGA from Triticum vulgaris ), N -acetylglucosamine, and fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit immunoglobulin were from Sigma; PVDF sheets were from Millipore; Affi-Gel 10 and nonfat dry milk were from Bio-Rad; HOE140 was from Hoechst; the TNT reticulolysate kit was from Promega; the first strand cDNA synthesis kit was from Pharmacia; Vent(exo+)DNA polymerase was from New England Biolabs; Centricon 30 filters were from Amicon. All other chemicals were of analytical grade.…”

Section: Methodsmentioning

confidence: 99%

Structure of the Bradykinin B₂Receptors' Amino Terminus

AbdAlla¹,

Godovac‐Zimmermann²,

Braun³

et al. 1996

Biochemistry

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The peptide hormone bradykinin exerts important biological functions by binding to and activating bradykinin B2 receptors. B2 receptors belong to the seven transmembrane domain (7TM) receptor family. Cloning of the cDNA sequences for the rat, human, and mouse bradykinin B2 receptor revealed several in-frame AUG triplets as potential initiation sites for translation. Due to "Kozak-like" consensus nucleotides, the AUG codon closest to transmembrane domain 1 was assumed the preferred initiation site for translation, but the real amino terminus of the B2 receptor protein was unknown. The amino terminus of several 7TM receptors has been shown to be essentially involved in receptor activation and/or ligand binding. Therefore we determined the amino terminus of the human and of the rat B2 receptor using domain-specific antipeptide antibodies, amino acid sequence analysis, and in vitro transcription/translation. We report that the human and rat B2 receptor protein start with the methionine which is translated from the first in-frame AUG. This start site extends the known amino terminus of the human and rat B2 receptors by 27 or 30 amino acid residues, respectively. Antibodies raised against a peptide of the initial 27 amino acids of the human B2 receptor stained B2 receptors on intact cells. This finding excludes the existence of a signal sequence for this receptor.

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“…Demonstration of analysis by 2D electrophoresis of transferrin receptor [7] or ACTH receptor [8] in complex membrane samples was obtained by immunoblotting. Conversely, analysis by 2D electrophoresis of semi-purified membrane preparations allowed to identify some membrane receptors by classical staining [9][10][11]. But in a few cases, definitive evidence of poor performance of classical 2D electrophoresis protocols with well-known membrane proteins could also be established [12], and the overall situation of the performance of 2D electrophoresis for membrane proteins using these classical protocols based on urea-detergent as the solubilization agent in IEF has been reviewed [13].…”

Section: A Historical Introduction: the Protoproteomics Eramentioning

confidence: 99%

Fully denaturing two‐dimensional electrophoresis of membrane proteins: A critical update

et al. 2008

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The quality and ease of proteomics analysis depends on the performance of the analytical tools used, and thus of the performances of the protein separation tools used to deconvolute complex protein samples. Among protein samples, membrane proteins are one of the most difficult sample classes, because of their hydrophobicity and embedment in the lipid bilayers. This review deals with the recent progresses and advances made in the separation of membrane proteins by 2-DE separating only denatured proteins. Traditional 2-D methods, i.e., methods using IEF in the first dimension are compared to methods using only zone electrophoresis in both dimensions, i.e., electrophoresis in the presence of cationic or anionic detergents. The overall performances and fields of application of both types of method is critically examined, as are future prospects for this field.

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Structural features of the human bradykinin B2 receptor probed by agonists, antagonists, and anti-idiotypic antibodies

Cited by 66 publications

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Na⁺ Ions Binding to the Bradykinin B₂ Receptor Suppress Agonist-Independent Receptor Activation

Na⁺ Ions Binding to the Bradykinin B₂ Receptor Suppress Agonist-Independent Receptor Activation

Structure of the Bradykinin B₂Receptors' Amino Terminus

Fully denaturing two‐dimensional electrophoresis of membrane proteins: A critical update

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Structural features of the human bradykinin B2 receptor probed by agonists, antagonists, and anti-idiotypic antibodies

Cited by 66 publications

References 0 publications

Na+ Ions Binding to the Bradykinin B2 Receptor Suppress Agonist-Independent Receptor Activation

Na+ Ions Binding to the Bradykinin B2 Receptor Suppress Agonist-Independent Receptor Activation

Structure of the Bradykinin B2Receptors' Amino Terminus

Fully denaturing two‐dimensional electrophoresis of membrane proteins: A critical update

Contact Info

Product

Resources

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Na⁺ Ions Binding to the Bradykinin B₂ Receptor Suppress Agonist-Independent Receptor Activation

Na⁺ Ions Binding to the Bradykinin B₂ Receptor Suppress Agonist-Independent Receptor Activation

Structure of the Bradykinin B₂Receptors' Amino Terminus